Separation of DNA fragments by high-resolution ion-exchange chromatography on a nonporous QA column.
A nonporous QA column (a strong anion exchanger) was used for HPLC of DNA fragments. This column was successfully employed to separate small (ca. 10 bp) and intermediate size (ca. 10 kbp) DNA fragments from each other. The column also separated double-stranded DNA from its single-stranded form, and circular DNA molecules from linear ones. The entire separation process was completed within 60 min. The recovery of DNA fragments in each run was above 95%. High resolution was obtained both at an analytical level (microgram scale) and at a preparative level (100 micrograms scale). In view of time efficiency, recovery, and resolution, the nonporous QA column is superior to other porous ion-exchange columns and expected to be a very useful tool in molecular biological studies.
Ohmiya
Y
Y
Department of Physical Biochemistry, Gunma University, Maebashi, Japan.
Kondo
Y
Y
Kondo
T
T
D000838:Anions; D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004247:DNA / Q000302:isolation & purification*; D004270:DNA, Circular / Q000302:isolation & purification; D004278:DNA, Superhelical / Q000302:isolation & purification; D004983:Ethanolamines; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D015203:Reproducibility of Results; D012965:Sodium Chloride
D000838:Anions; D002021:Buffers; D004270:DNA, Circular; D004278:DNA, Superhelical; D004983:Ethanolamines; D012965:Sodium Chloride; D004247:DNA; C007369:2-diethylaminoethanol